Accurate and precise real-time RT-PCR assays for the identification of astrovirus associated encephalitis in cattle

Lüthi, Ramona; Boujon, Céline L.; Kauer, Ronja; Koch, Michel C.; Bouzalas, Ilias G.; Seuberlich, Torsten (2018). Accurate and precise real-time RT-PCR assays for the identification of astrovirus associated encephalitis in cattle. Scientific Reports, 8(1), p. 9215. Nature Publishing Group 10.1038/s41598-018-27533-8

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A novel bovine astrovirus genotype species (BoAstV-CH13/NeuroS1) was recently identified in brain tissues of cattle as a plausible cause of encephalitis. The purpose of the present study was to develop and validate real time RT-PCR assays for the detection of BoAstV-CH13/NeuroS1 in brain tissues of cattle. Three different primer-probe combinations were designed based on BoAstV-CH13/NeuroS1 full-genome sequences of 11 different strains identified in cattle, and established in three distinct one-step real time RT-PCR protocols. These protocols were compared regarding their diagnostic performance using brain tissues of cattle with and without astrovirus associated encephalitis. The limit of detection (LOD) of all three assays was between 1.34 × 101 and 1.34 × 102 RNA copies, leading to an analytical sensitivity two orders of magnitude superior compared to a conventional pan-astrovirus RT-PCR protocol (LOD 1.31 × 104 RNA copies). Amplification efficiency was in the range of 97.3% to 107.5% with linearity (R2) > 0.99. The diagnostic sensitivity and specificity of the assays was determined as 100%, and all three revealed good intra- and inter-test repeatability. In conclusion, the newly developed RT-qPCRs are sensitive, specific, and reliable test formats that will facilitate BoAstV-CH13/NeuroS1 detection in routine diagnostics as well as in research settings.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Research Foci > NeuroCenter
05 Veterinary Medicine > Department of Clinical Research and Veterinary Public Health (DCR-VPH) > Experimental Clinical Research
05 Veterinary Medicine > Department of Clinical Research and Veterinary Public Health (DCR-VPH)

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

Lüthi, Ramona; Boujon, Céline Louise; Kauer, Ronja Veronique; Koch, Michel Christoph; Bouzalas, Ilias and Seuberlich, Torsten

Subjects:

500 Science > 570 Life sciences; biology
500 Science > 590 Animals (Zoology)
600 Technology > 630 Agriculture

ISSN:

2045-2322

Publisher:

Nature Publishing Group

Language:

English

Submitter:

Céline Boujon

Date Deposited:

20 Sep 2019 14:47

Last Modified:

30 Oct 2019 07:43

Publisher DOI:

10.1038/s41598-018-27533-8

PubMed ID:

29907784

BORIS DOI:

10.7892/boris.130513

URI:

https://boris.unibe.ch/id/eprint/130513

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